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. 2014 Aug 26;42(16):10720–10730. doi: 10.1093/nar/gku778

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© The Author(s) 2014. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 4. — AUF1 binds to FST mRNA and dissociates during glucose deprivation. (A) GST and GST-AUF1 isoforms were expressed in E. coli, puriﬁed and analysed by Coomassie blue staining. (B) FST 3′UTR RNA was incubated with increasing amounts of the indicated proteins. RNA–protein interaction was evaluated by RNA gel shift experiments. (C) Pull-down assay was performed using biotinylated FST RNA fragments and HeLa whole-cell lysates followed by immunoblotting. (D, E, F) Immunoprecipitation was performed in cells cultured with or without glucose using anti-AUF1 antibody or normal immunoglobulin G (IgG). FST mRNA (D), p21 mRNA (E) or Cyclin D1 mRNA (F) was determined by RT-qPCR.